표제지
목차
국문초록 12
1. 서론 14
가. 온라인 모니터링 기술 14
1) 흐름주입 분석법(flow injection analysis, FIA) 14
2) 흐름분석법의 원리 및 장치구성 15
3) 분할흐름분석법(Segmented Flow Analysis, SFA) 15
4) 순차주입분석법(sequential injection analysis, SIA) 16
2. 재료 및 방법 18
가. 효소 및 시약 18
1) 젖산 18
2) 숙신산 19
3) 푸마르산 20
나. 측정원리 21
1) 젖산 21
2) 숙신산 21
3) 푸마르산 22
다. 효소(LOD, LDH, ICL/ICDH, Fum/MDH)의 고정화 22
라. FIA 장치 구성 및 운전 24
1) 젖산-FIA 장치 24
2) 숙신산-FIA 및 푸마르산 FIA 장치 24
마. 오프라인 분석 30
1) 젖산의 분석 30
2) 숙신산의 분석 30
3) 푸마르산의 분석 30
3. 결과 및 고찰 31
가. 고정화 효소의 특성 31
1) 젖산-FIA 31
2) 숙신산-FIA 48
3) 푸마르산-FIA 61
나. 생물공정의 모니터링 74
1) 젖산의 모니터링 74
2) 숙신산의 모니터링 76
3) 푸마르산 모니터링 80
4. 요약 84
가. 젖산-FIA 84
나. 숙신산-FIA 85
다. 푸마르산-FIA 85
5. 참고문헌 86
Abstract 89
Table 1. Reference operating conditions for a LOD-FIA and LDH-FIA system. 27
Table 2. Reference operating conditions of the Succinate-FIA system 28
Table 3. Reference operating conditions for a Fumarate-FIA system. 29
Figure 1. Comparison of FIA and SIA system and dispersion of sample. 17
Figure 2. Schematic of a LOD/LDH-FIA system. I; injector, IER; Immobilized... 25
Figure 3. Schematic of a Succinate/Fumarate-FIA system. D; fluorescence... 26
Figure 4. Linear detection range of a LOD-FIA system under the operating... 33
Figure 5. Effects of carrier solution flow rates on the activity of immobilized LOD. 34
Figure 6. Effects of carrier solution pH values on the activity of immobilized LOD. 35
Figure 7. Effects of reaction temperature on the activity of immobilized LOD 36
Figure 8. Relative peak heights of 0.5 g/L L-lactate in absence and presence of... 37
Figure 9. Relative peak heights of 0.5 g/L L-lactate in absence and presence of... 38
Figure 10. Linear detection range of a LDH-FIA system under the operating... 41
Figure 11. Effects of carrier solution flow rates on the activity of immobilized LDH. 42
Figure 12. Effects of carrier solution pH on the activity of immobilized LDH 43
Figure 13. Effects of NAD+ injection flow rates on the activity of immobilized LDH[이미지참조] 44
Figure 14. Relative peak heights of 5 g/L L-lactate at different sample injection volumes. 45
Figure 15. Relative peak heights of 5 g/L L-lactate in absence and presence of... 46
Figure 16. Relative peak heights of 5 g/L L-lactate in absence and presence of... 47
Figure 17. Operational stability of the immobilized isocitrate lyase and isocitrate... 50
Figure 18. Effects of pH values on the activity of the immobilized isocitrate... 51
Figure 19. Effects of sample injection volume on the peak heights of the FIA... 52
Figure 20. Effects of carrier buffer flow rates on the peak heights of the FIA... 53
Figure 21. Effects of NADP+ concentrations on the peak heights of the FIA...[이미지참조] 54
Figure 22. Effects of NADP+ solution flow rates on the peak heights of the FIA...[이미지참조] 55
Figure 23. Effects of reaction temperature on the peak heights of the FIA... 56
Figure 24. Effects of salts contained in sample on the activity of the... 57
Figure 25. Effects of metabolites contained in sample on the activity of the... 58
Figure 26. Effects of substrates contained in sample on the activity of the... 59
Figure 27. Influence of some components added to carrier buffer solution on the... 60
Figure 28. Operational stability of the immobilized fumarase and malic... 64
Figure 29. Effects of pH values on the activity of the immobilized fumarase and... 65
Figure 30. Effects of sample injection volume on the peak heights of the FIA... 66
Figure 31. Effects of carrier buffer flow rates on the peak heights of the FIA... 67
Figure 32. Effects of NAD+ concentrations on the peak heights of the FIA...[이미지참조] 68
Figure 33. Effects of NADP+ solution flow rates on the peak heights of the FIA...[이미지참조] 69
Figure 34. Effects of reaction temperature on the peak heights of the FIA... 70
Figure 35. Effects of metabolites contained in sample on the activity of the... 71
Figure 36. Effects of nutrient salts and substrates contained in sample on the... 72
Figure 37. Influence of some components added to carrier buffer solution on the... 73
Figure 38. On-line monitoring of L-lactate in a simulated bioprocess by a...[이미지참조] 75
Figure 39. Linear concentration range of succinate by the sucinate-FIA system. 77
Figure 40. On-line monitoring of succinate in a simulated bioprocess by a... 78
Figure 41. On-line monitoring of the concentrations of succinate during a... 79
Figure 42. Effects of the packing ratio of two immobilized fumarase and malric... 81
Figure 43. On-line monitoring of the concentrations of fumaric acid in a... 82
Figure 44. On-line monitoring of the concentrations of fumaric acid during a... 83